GD2-CAR T Cells for Pediatric Brain Tumours

Inclusion Criteria:

1. Imaging assessments performed within 14 days of start of treatment

2. Age: 6months-30years

3. Measurable or evaluable disease on at least 2 dimensions on MRI at the time oftreatment enrollment

4. Karnofsky/Lansky≥60

5. Recoverfromthetoxiceffectsofpreviousradiationandchemotherapies:grade4and or 3non-hematologic toxicities must have resolved to grade ≤ 2; in presence of chroniccomplications (i.e. treatment-associated thrombocytopenia), patient must beclinically stable, according to the opinion of the treating physicians, and meet allother eligibility criteria

6. Positioning of an implantable intraventricular access device (CodmanHolterRickhamreservoir, Integra LifeSciences, NJ, U.S.A) and a microdialysis probe (71 highcutoff microdialysis bolt catheter, M Dialysis AB, Stockholm Sweden)

7. Written and signed informed consent from patients, parents or legal guardians. Forsubjects < 18 year-old their legal guardian must give informed consent. In addition,pediatric subjects will be included in age-appropriate discussion and writteninformed assent will be obtained for those greater than or equal to 7 years of age,when appropriate

8. Patients of childbearing or child-fathering potential must be willing to practicebirth control from the time of enrollment on this study and for four months afterreceiving the preparative regimen

9. Females of childbearing potential must have a negative pregnancy test because of thepotentially dangerous effects on the fetus

Exclusion Criteria:

1. Pregnant or lactating women

2. Severe,uncontrolledactiveinfections

3. HIV or active HCV and/or HBV infection

4. Rapidly progressive disease with life expectancy < 6 weeks

5. Historyofgrade3or4hypersensitivitytomurineprotein-containingproducts

6. Hepatic function: inadequate liver function defined as total bilirubin > 4x upperlimit of normal (ULN) or transaminase (ALT and AST) > 6 x ULN based on age andlaboratory specific normal ranges

7. Renal function: serum creatinine > 3x ULN for age

8. Blood oxygen saturation < 90%

9. Cardiac function: left ventricular ejection fraction lower than 45% by ECHO

10. Marrow function: absolute neutrophils count (ANC) lower than 500/mm3 and/orplatelets lower than 20.000 (not reached by transfusion)

11. Congestive heart failure, cardiac arrhythmia, psychiatric illness, or socialsituations that would limit compliance with study requirements or in the opinion ofthe principal investigator (PI) would pose an unacceptable risk to the subject. 12.Concurrent or recent prior therapies, before infusion:

12. If receiving glucocorticoids, patient must be on a stable or weaning dose forat least 7 days prior to infusion. Recent or current use ofinhaled/topical/non- absorbable steroids is not exclusionary. Subjectsreceiving steroid therapy at physiologic replacement doses only are allowedprovided there has been no increase in dose for at least 2 weeks prior tostarting apheresis

13. Systemic chemotherapy in the 3 weeks preceding infusion

14. Immunosuppressive agents less than or equal to 30 days

15. Radiation therapy must have been completed at least 6 weeks prior to enrollment

16. Otheranti-neoplasticinvestigationalagentscurrentlyorwithin30dayspriorto startof protocol therapy

13.Patient-derived GD2-CART01 production failure: vitality <80%, CD3+ cells <80%, CD3+ CAR+ cells <20%, CD3+ CAR+ antitumor activity <60% in functional co-culture assay at an Effector: Target ratio 1:1, viable CAR+ cells upon AP1903 exposition >20%, RCR positivity, Vector Copy Number >10, non-sterility, endotoxin contamination (> 1 EU/ml)